Integrative structural biology uses a large variety of biophysical methods that provide detailed knowledge of the atomic and molecular structure of macromolecules, isolated or in their native environment. This provides a better understanding of the basic of chemistry of life and helps with the development of new biotechnology tools, drugs, and biomaterials. Access to large-scale installations (synchrotrons producing X-rays, state-of-the-art electron microscopes, powerful NMR magnets) is necessary for research in structural biology. This Methods Collection will describe the most important methods for the structural analysis of macromolecules, applications in fragment-based lead drug discovery, methods to investigate the structure of macromolecules in their native environment, and high throughput approaches for structural determination. It will consist of a variety of methods that includes, but is not limited to, NMR, cryo-EM, X-ray crystallography, and scattering.